Gene Editing of the Jasmonate Biosynthesis OsAOC Gene in Rice

A procedure to develop new Jasmonate (JA)-deficient mutant using CRISPR-Cas9 in rice was presented by the researchers from Universite de Montpellier, France and partners in Springer Protocols. They targeted the Allene Oxide Cyclase (OsAOC) gene because it is a single copy gene in the JA biosynthesis pathway in rice.

The role of JA in various plant physiological processes and its interactions with the environment has been well-documented. Mutants with dysfunctional JA production and/or signaling are useful in investigating the function of JA, which is a phytohormone. However, previous studies presented mutants that were mostly generated using random mutagenesis. In the recent study, CRISPR-Cas9 was employed for a more precise genome editing at specific loci while limiting unwanted mutations.

The protocol presented describes the selection of the 20-nt target sequence, construction of the binary vector, and strategy for selecting the T-DNA-free mutant.

Read more from the book Methods in Molecular Biology.